藏藥臭蚤草中 11個倍半萜化學成分研究

黃圣卓,蔣思萍,朱華結

1中國科學院昆明植物研究所植物化學與植物資源持續利用國家重點實驗室,昆明 650204;2西藏高原生物研究所,拉薩 850001;3中國科學院研究生院,北京 100049

藏藥臭蚤草中 11個倍半萜化學成分研究

黃圣卓1,3,蔣思萍2,朱華結1＊

1中國科學院昆明植物研究所植物化學與植物資源持續利用國家重點實驗室,昆明 650204;2西藏高原生物研究所,拉薩 850001;3中國科學院研究生院,北京 100049

首次進行中國特有菊科藏藥臭蚤草 (Pulicaria insignisDrumm.ex Dunn)的化學成分研究,從全草中分離得到 11個倍半萜類化合物,通過波譜數據分析分別鑒定為 Xanthanolide(1),(3αR,4αR,5R,7αS,9αS)-5-Hydroxy-5-methyl-3,8-dimethylenedecahydroazuleno[6,5-β]furan-2(3H)-one(2)10α-Hydroxy-14H-inuviscolide(3), (3R,3αR,4αS,7αS,8S,9αS)-3,4α,8-Tr imethyldecahydroazuleno[6,5-β]furan-2,5-dione(4)8-Epi-conferti(5), (3αR,5S,11αS,E)-5-Hydroxy-10-methyl-3,6-dimethylene-3α,4,5,6,7,8,11,11α-octahydrocyclodeca[α]furan-2 (3H)-one(6),(3αR,8αR,9αR)-5,8α-Dimethyl-3-methylene-3α,4,8,8α,9,9α-hexahydronaphtho[2,3-β]furan-2,6 (3H,7H)-dione(7),Pterodonoic acid(8),8-Epi-isovangustin(9),(3αR,4αS,8R,8αR,9αR)-8-Hydroxy-5,8α-dimethyl-3-methylene-3,3α,4,4α,8,8α,9,9α-octahydronaphtho[2,3-β]furan-2(7H)-one(10),(5S,8αR,9αR)-3-(Hydroxymethyl)-5,8α-dimethyl-6,7,8,8α,9,9α-hexahydronaphtho[2,3-β]furan-2(5H)-one(11)。所有化合物均首次從該植物中分離得到,同時補充了文獻中化合物 6和 9所缺少的碳譜數據。

藏藥;臭蚤草;倍半萜

臭蚤草Pulicaria insignisDrumm.ex Dunn是菊科 Compositea(Asteraceae)旋復花族Inluneae蚤草屬Pulicaria的多年生草本。僅產西藏南部江孜、拉薩、察日壩、沙伽、尼木、日喀則、南太林等處。生于山脊巖石上,常見于石礫坡地和草叢中,海拔 4000～4310 m,根狀莖粗壯、多分枝,“成片叢生,植物有粘液,具惡臭”[1],為西藏特有的藥用植物。臭蚤草的花在西藏供藥用,藏名“Ming·chen·serpo”;全草味苦性涼,具有鎮咳舒肝,清血熱,透骨蒸。用于肺癆咳嗽,兩肋疼痛,勞熱骨蒸,花序有清熱、止痛的功效,還有驅跳蚤的作用[2]。目前人們從該屬植物中分離到的化合物主要為倍半萜、二萜和黃酮等化合物[3-10],未見臭蚤草的化學成分報道。為了了解臭蚤草的活性成分,我們對該植物全草進行化學成分研究,從中分離得到 11化合物,所有化合物均首次從該植物中分離得到,同時補充了文獻中化合物6和 9所缺少的碳譜數據。

1 儀器和材料

VGA Autospec-3000型質譜儀;Bruker AM-400和DRX-500超導核磁共振儀,T MS(四甲基硅烷)為內標;JASCO D IP-370和 OA AA-55型數字旋光儀,單位為 c單位為 g/mL;薄層色譜和正相柱色譜硅膠(青島海洋化工);Sephadex LH-20(Phar macia公司);ODS(Y MC公司);MCI樹脂(三菱公司)。

植物樣品于 2007年 10月采于西藏拉薩,植物標本由西藏高原生物所蔣思萍研究員鑒定為Pulicaria insignisDrumm.exDunn,標本保存于西藏高原生物所標本室。

2 提取和分離

90 Kg干燥臭蚤草全草,粉碎后用甲醇提取濃縮獲得浸膏,石油醚脫脂后用乙酸乙酯萃取 10次濃縮獲得浸膏 780 g。乙酸乙酯部石油醚 /乙酸乙酯(20∶1-0∶1)分為 5個部分 (20∶1,10∶1,5∶1,2∶1, 0∶1),其中石油醚/乙酸乙酯 (10∶1)部分經MCI樹脂脫色(收集甲醇洗脫部分),再經石油醚/丙酮 (50∶1-10∶1)和氯仿/丙酮 (100∶1-20∶1)多次反復梯度洗脫,經ODS柱(甲醇/水 1∶4-4∶1)梯度洗脫后,再分別經 SephadexLH-20凝膠(氯仿/甲醇 1∶1洗脫)純化,分別獲得化合物 1(256 mg),2(11.1 mg),3(851 mg),4(16.2 mg),5(382 mg),6(8.8 mg),7(20 mg),8 (8.8 mg),9(171 mg),10(4 mg),11(18.8 mg)。

3 結構鑒定

Xanthanolide(1) 白色塊狀晶體,C15H20O3; mp.128～129℃,+26.6(c0.0064 g/mL,下同,MeOH);1H NMR(CDCl3,500 MHz)δ(ppm): 6.12(d,J=3.2 Hz,1H),5.43(d,J=3.2 Hz,1 H),5.33(m,1H),4.54(td,J=8.7,2.7 Hz,1H), 3.22(m,1H),2.39(m,1H),2.30(m,3H),2.10 (m,3 H),2.08(s,3H),1.87(m,1H),1.80(dd, 1H),1.02(d,J=7.0 Hz,3H),ESI-MS(positive)m/z:249[M+1]+,271[M+Na]+,228;碳譜數據見表 1。其波譜數據與文獻[11]報道一致。

(3αR,4αR,5R,7αS,9αS)-5-Hydroxy-5-methyl-3,8-d imethylenedecahydroazuleno[6,5-β]furan-2(3H)-one(2) 白色塊狀晶體,C15H20O3;mp.132～133℃;+62.3(c0.0014,CHCl3);1H NMR (CDCl3,400 MHz)δ(ppm):2.69(1H,d,J=10.5 Hz),1.96(1H,m),1.65(1H,m),1.85(m,1 H), 1.65(m,1H),1.71(1H,m),2.28(1H,t,J= 13.3,3.7 Hz),1.24(1H,m),2.66(1H,m),4.31 (1H,ddd,J=10.6,9.3,1.6 Hz),2.55(1H,dd,J= 15.5,10.6 Hz),6.22(1H,d,J=3.5 Hz),5.54 (1H,d,J=3.5 Hz),5.09(1H,s),4.97(1H,s), 1.20(3H,s),EI-MSm/z:248[M]+,230[MH2O]+,215,190,145;碳譜數據見表 1。其波譜數據與文獻[12]報道一致。

10α-Hydroxy-14H-inuviscolide(3) 無色油狀物質,C15H22O4;+56.7(c0.00335 g/mL, CHCl3);1H NMR(CDCl3,500 MHz)δ(ppm):6.00 (1H,d,J=3.5 Hz),5.40(1H,d,J=3.5 Hz),4.25 (1H,dd,J=12.5,10.0 Hz),2.43(1H,ddd,J= 11.0,10.5,3.5 Hz),2.33(1H,d,J=14.5 Hz), 2.20(1H,ddd,J=12.5,4.0,2.0 Hz),1.89(1H, dd,J=14.5,12.0 Hz),1.83(1H,m),1.67(1H, m),1.54(2H,m),1.48(1H,ddd,J=11.5,11.0, 2.0 Hz),1.67(3H,s),1.05(3H,s),EI-MSm/z: 267[M+1]+,230[M-H2O]+,191,190,95,81;碳譜數據見表 1。其波譜數據與文獻[12]報道一致。

(3R,3αR,4αS,7αS,8S,9αS)-3,4α,8-Tr imethyldecahydroazuleno[6,5-β]furan-2,5-dione(4)無色油狀物質,C15H22O3;+59.2(c0.0071 g/mL,CHCl3),1H-NMR (CDCl3,400 MHz)δ (ppm):1.81(1H,m),2.01(1H,m),2.05(1H, m),2.20(1H,ddd,J=19.0,9.0,9.0 Hz),2.49 (1H,ddd,J=19.0,9.0,2.0 Hz),2.57(1H,dd,J= 15.0,5.0 Hz),1.30(1H,dd,J=15.0,11.5 Hz), 1.85(1H,ddd,J=11.5,11.0,9.0 Hz),4.29(1H, ddd,J=12.0,9.0,3.0 Hz),2.29(1H,m),1.74 (1H,ddd,J=12.0,12.0,6.0 Hz),2.31(1H,m), 2.33(1H,m),1.22(3H,d,J=7.0 Hz),1.18(3H, d,J=7.0 Hz),1.08(3H,s),FAB-MS(positive)m/ z:251[M+1]+,233,111;碳譜數據見表 1。其波譜數據與文獻[13]報道一致。

8-Epi-confertin(5) 白色塊狀晶體,C15H20O3;mp.125～126℃,+163.5(c0.00315 g/mL,CHCl3),1H NMR(CDCl3,400 MHz)δ(ppm): 6.18(1H,d,J=3.5 Hz),5.53(1H,d,J=3.5 Hz), 4.28(1H,td,J=8.8,2.4 Hz),2.73(1H,m),2.67 (1H,dd,J=14.8,5.3 Hz),2.50(1H,m),2.30 (2H,m)2.17(1H,dd,J=19.5,9.7 Hz),2.15 (1H,m),2.08(1H,m),1.85(2H,m),1.43(1H, t,J=11.8 Hz),1.16(3H,d,J=7.6 Hz,),1.07 (3H,s),EI-MSm/z:248[M]+,228,230,215,204, 189,136,81;碳譜數據見表 1。其波譜數據與文獻[12]報道一致。

(3αR,5S,11αS,E)-5-Hydroxy-10-methyl-3,6-d imethylene-3α,4,5,6,7,8,11,11α-octahydrocyclodeca[β]furan-2(3H)-one(6) 淡黃色油狀物質,C15H20O3;-52.63 (c0.00152 g/mL, MeOH),1H NMR(CDCl3,400 MHz)δ(ppm):6.30 (1H,d,J=3.0 Hz),6.18(1H,d,J=3.0 Hz),5.50 (1H,dd,J=7.7,7.5 Hz),5.33(1H,s),5.16(1H, s),4.14(1H,m),4.08(1H,d,J=6.3 Hz),2.75 (2H,m),2.38(2H,m),2.04(1H,t,J=11.6 Hz), 1.96(1H,dd,J=7.0,14.5 Hz),1.85(1H,m), 1.73(2H,m),1.70(3H,s),ESI-MS(positive)m/ z:249[M+1]+,271[M+Na]+;碳譜數據見表 1。其波譜數據與文獻[14]報道一致。

(3αR,8αR,9αR)-5,8α-D imethyl-3-methylene-3α,4,8,8α,9,9α-hexahydronaphtho[2,3-β]furan-2,6(3H,7H)-dione(7) 白色粉末,C15H18O3;+17.3(c0.00075 g/mL,MeOH),1H NMR (CDCl3,500 MHz)δ(ppm):6.38(1H,d,J=3.2 Hz),5.72(1H,d,J=2.6 Hz),4.61(1H,ddd,J= 11.5,8.5,4.7 Hz),3.22(1H,m),3.04(1H,dd,J=13.1,7.3 Hz),2.60(1H,td,J=14.1,5.5 Hz), 2.46(1H,dt,J=14.1,3.1 Hz),2.23(1H,t,J= 14.5 Hz),2.05(1H,m),1.94(1H,dd,J=13.8, 4.7 Hz),1.83(3H,s),1.60-1.79(2H,m),1.24 (3H,s),FAB-MS(positive)m/z:247[M+1],154, 136,107,83;碳譜數據見表 1。其波譜數據與文獻[15]報道一致。

Pterodonoic acid(8) 白色粉末,C15H20O3;+17.3(c0.00075 g/mL,CH3OH)1H NMR (CDCl3,500 MHz)δ (ppm):6.41(1H,s),5.76 (1H,s),2.88(1H,d,J=14.5 Hz),2.41-2.84 (3H,m),2.06(1H,t,J=13.6 Hz),1.70-1.85 (5H,m),1.79(s,3H),1.51(2H,dt,J=4.1,12.7 Hz),1.25(3 H,s),ESI-MS(positive)m/z:249[M +1]+,271[M+K]+,519[2 M+Na]+;碳譜數據見表 1。其波譜數據與文獻[16]報道一致。

8-Epi-isovangustin(9) 白色塊狀晶體,C15H24O3;mp.138～140℃,+17.3(c0.0035 g/ mL,CH3OH),1H NMR(CDCl3,400MHz)δ(ppm): 6.13(1H,d,J=1.1 Hz),5.63(1H,d,J=1.1 Hz), 2.46(1H,tt,J=8.5,3.5 Hz),1.94(1H,d,J=12.4 Hz),1.70(1H,dd,J=12.4 Hz),1.70～1.15 (11H,m),1.07(3H,s),0.93(3H,s),ESI-MS (positive)m/z:251[M+Na]+,503[2M+1]+;碳譜數據見表 1。其波譜數據與文獻[17]報道一致。

(3αR,4αS,8R,8αR,9αR)-8-Hydroxy-5,8αd imethyl-3-methylene-3,3α,4,4α,8,8α,9,9α-octahydronaphtho[2,3-β]furan-2(7H)-one(10) 無色油狀物質,C15H20O3;[α]21D-31.5(c0.0016 g/mL, MeOH),1H NMR(CDCl3,400 MHz)δ(ppm):3.7 (1H,dd,J=10.0,7.0 Hz),4.05(1H,ddd,J= 12.5,12.5,4.0 Hz),5.34(1H,s),6.09(1H,d,J= 3.0 Hz),5.45(1H,d,J=3 Hz),2.51(1H,dd,J= 12.5,4.0 Hz),2.30-2.40(2H,m),2.25(1H,ddd,J=12.5,12.5,3.0 Hz),2.17(1H,d,J=3.0 Hz), 2.01(1H,dddq,J=12.0,10.0,2.15,2.15 Hz), 1.69(3H,s),1.42(2H,m),0.89(3H,s),ESI-MS (positive)m/z:249[M+1]+,271[M+Na]+,519 [2M+Na]+,131,185,157,119,105;碳譜數據見表1。其波譜數據與文獻報道一致[18].

(5S,8αR,9αR)-3-(Hydroxymethyl)-5,8αd imethyl-6,7,8,8α,9,9α-hexahydronaphtho[2,3-β]furan-2(5H)-one(11) 淡黃色油狀物質,C15H20O3;+249.2(c0.00915 g/mL,CHCl3);1H NMR(CDCl3,400 MHz)δ(ppm):1.60(1H,m), 1.68(1H,dd,J=13.0,4.0 Hz),1.56(1H,m), 1.95(1H,m),1.56(1H,m),1.75(1H,m),2.75 (1H,ddq,J=7.5,6.5,2.0 Hz),6.36(1H,s),4.78 (1H,dd,J=13.0,6.0 Hz),2.14(1H,dd,J=13.0, 6.0 Hz),1.54(1H,dd,J=13.0,13.0 Hz),4.40 (2H,s),1.32(3H,s),1.29(3H,d,J=7.5 Hz), 2.68(br),EI-MSm/z:248[M]+,230[M-H2O]+, 215,159,146;HR-ES IMS(positive)[M+Na]+271.1314,Calcd for[C15H20O3+Na]+m/z: 271.1310;碳譜數據見表 1。其波譜數據與文獻報道一致[19].

圖 1 臭蚤草中分離到的化合物 1～11Fig.1 Compounds 1-11 isolated fromPulicaria insignis

表 1 化合物 1～11的13C NMR數據 (13C,100 MHz in CDCl3)δ(ppm)Table 113C NMR of compounds 1-11(13C,100 MHz in CDCl3)δ(ppm).

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Sesquiterpenoids from Tibetan Folk DrugPulicaria insignis

HUANG Sheng-zhuo1,3,J IANG Si-ping2,ZHU Hua-jie1＊

1State key laboratory of Phytochem istry and Plant Rescources of W est of China,Kunm ing Institute of Botany,Academy of Sciences Kunm ing 650204,Yunnan China;2Plateau Institute of B iology Lhasa 850001,Tibet China;3Graduate University of Chinese Academy of Sciences Beijing 100049,China

Eleven known sesquiterpenoids were isolated from the Tibetan Folk DrugPulicaria insignis.On the basis of MS and NMR data,theywere identified as Xanthanolide(1),(3αR,4αR,5R,7αS,9αS)-5-Hydroxy-5-methyl-3,8-d imethylenedecahydroazuleno[6,5-β]furan-2(3H)-one(2),10α-hydroxy-14H-inuviscolide(3),(3R,3αR,4αS,7αS,8S,9αS)-3,4α,8-Trimethyldecahydroazuleno[6,5-β]furan-2,5-dione(4),8-Epi-conferti(5),(3αR,5S,11αS,E)-5-Hydroxy-10-methyl-3,6-dimethylene-3α,4,5,6,7,8,11,11α,-octahydrocyclodeca[α]furan-2(3H)-one(6),(3αR,8αR,9αR)-5,8α-Dimethyl-3-methylene-3α,4,8,8α,9,9α-hexahydronaphtho[2,3-β]furan-2,6(3H,7H)-dione(7), Pterodonoic acid(8),8-Epi-isovangustin(9),(3αR,4αS,8R,8αR,9αR)-8-Hydroxy-5,8α-d imethyl-3-methylene-3, 3α,4,4α,8,8α,9,9α-octahydronaphtho[2,3-β]furan-2(7H)-one(10)(5S,8αR,9αR)-3-(Hydroxymethyl)-5,8αd imethyl-6,7,8,8α,9,9α-hexahydronaphtho[2,3-β]furan-2(5H)one(11).All compounds were isolated from this plant for the first t ime,and the13C NMR data of compounds 6 and 9 were shawed in this paper.

Tibetan drug;Pulicaria insignis;chemical constituents;sesquiterpenoids

R284.2;Q946

A

1001-6880(2010)05-0736-05

2009-04-30 接受日期:2009-10-20

國家自然科學基金(NSFC#30770235);中國科學院院地合作基金(YZ-06-1)

＊通訊作者 Tel:86-871-5216179;E-mail:hjzhu@mail.kib.ac.cn